Dietary supplement containing phospholipid-DHA derived from eggs

ABSTRACT

Described herein are manufactured dietary supplements that contain a phospholipid extract and the extract contains two forms of phospholipid-docosahexanoic acid. One form of phospholipid-docosahexanoic acid is phosphatidylcholine-docosahexanoic acid, and another form of phospholipid-docosahexanoic acid is phosphatidylethanolamine-docosahexanoic acid.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of nonprovisional U.S.application Ser. No. 14/562,650 filed Dec. 5, 2014, which claimspriority to Provisional U.S. Application No. 61/912,205 filed on Dec. 5,2013 and Provisional U.S. Application No. 62/088,528 filed on Dec. 5,2014, which are hereby incorporated by reference in its entirety.

FIELD

The present disclosure generally relates to dietary supplementsproviding phospholipid-docosahexanoic acid derived from eggs.

BACKGROUND

Polyunsaturated fatty acids have numerous crucial biological functionsin mammals. Moreover, in adult humans, certain polyunsaturated fattyacids have been linked to potentially promoting improved cognitiveability and have been theorized as potential treatments for arthritis,cancer, diabetes, lupus, and psoriasis. Also, certain polyunsaturatedfatty acids have been linked to promoting retinal and brain developmentin fetuses and newborns.

SUMMARY

A manufactured dietary supplement contains a phospholipid extractderived from eggs and the extract contains two forms ofphospholipid-docosahexanoic acid derived from eggs. One form ofphospholipid-docosahexanoic acid is phosphatidylcholine-docosahexanoicacid, and another form of phospholipid-docosahexanoic acid isphosphatidylethanolamine-docosahexanoic acid. And the phospholipidextract has less than about 3% by weight of free triglycerides.

DESCRIPTION OF THE FIGURES

These and other features, aspects and advantages of the presentdisclosure are better understood when the following detailed descriptionof the disclosure is read with reference to the accompanying drawing, inwhich:

FIG. 1 is a schematic depiction of docosahexanoic acid metabolism in asubject.

FIG. 2 is a schematic depiction of phosphatidylcholine-docosahexanoicacid.

FIG. 3 is a schematic depiction ofphosphatidylethanolamine-docosahexanoic acid.

DETAILED DESCRIPTION

Before the present compounds, compositions, and/or methods are disclosedand described, it is to be understood that the aspects described beloware not limited to specific compounds, synthetic methods, or uses assuch may, of course, vary. It is also to be understood that theterminology used herein is for the purpose of describing particularaspects only and is not intended to be limiting.

In this specification and in the claims that follow, reference will bemade to a number of terms that shall be defined to have the followingmeanings:

It must be noted that, as used in the specification and the appendedclaims, the singular forms “a,” “an” and “the” include plural referentsunless the context clearly dictates otherwise. Thus, for example,reference to “a microparticle” includes one or more microparticles.

“Optional” or “optionally” means that the subsequently described eventor circumstance may or may not occur, and that the description includesinstances where the event or circumstance occurs and instances where itdoes not.

As used herein, “subject” refers to a mammal, including a human that isin need of supplementation and/or benefits from the compositions andmethods described herein.

As used herein, “organic” refers to a food that complies with U.S.D.Aregulations set out herein below.

As used herein, the term “rounded” means that an outer periphery of astructure is substantially free of angularity. For example, the outerperiphery of microparticles can have spherical shapes and oval shapesthat are free of angularity and have rounded shapes.

As used herein, the term “about” is used to provide flexibility to anumerical range endpoint by providing that a given value may be“slightly above” or “slightly below” the endpoint without affecting thedesired result.

The disclosure provides various teachings on dietary supplementscontaining phospholipid extracts derived from eggs. The ingredients,compounds, chemicals that make up the elements of the dietarysupplements may contain vitamins, foods, or other organic matter that isprone to degradation after formulation in a finished dietary supplement.It is common practice to formulate a dietary supplement to include an“overage” or an additional amount of ingredient. Any amount ofingredient, compound, or chemicals that is within an “overage” of theranges and amounts claimed and exemplified in this disclosure would beconsidered equivalent.

The disclosure provides dietary supplement that contains a phospholipidextract that may be made up of different forms of phospholipids indifferent ratios and amounts. Where the disclosure refers to an amountof a specific phospholipid, it is measured by weight and not by molaramount. Where a phospholipid extract refers to a ratio between differentphospholipids, it is measured by weight and not by molar amount.

As used herein, a plurality of items, structural elements, compositionalelements, and/or materials may be presented in a common list forconvenience. However, these lists should be construed as though eachmember of the list is individually identified as a separate and uniquemember. Thus, no individual member of such list should be construed as ade facto equivalent of any other member of the same list solely based ontheir presentation in a common group without indications to thecontrary.

Concentrations, amounts, and other numerical data may be expressed orpresented herein in a range format. It is to be understood that such arange format is used merely for convenience and brevity and thus shouldbe interpreted flexibly to include not only the numerical valuesexplicitly recited as the limits of the range, but also to include allthe individual numerical values or sub-ranges encompassed within thatrange as if each numerical value and sub-range is explicitly recited. Asan illustration, a numerical range of “about 1 to about 5” should beinterpreted to include not only the explicitly recited values of about 1to about 5, but also include individual values and sub-ranges within theindicated range. Thus, included in this numerical range are individualvalues such as 2, 3, and 4 and sub-ranges such as from 1-3, from 2-4,and from 3-5, etc., as well as 1, 2, 3, 4, and 5, individually. Thissame principle applies to ranges reciting only one numerical value as aminimum or a maximum. Furthermore, such an interpretation should applyregardless of the breadth of the range or the characteristics beingdescribed.

The nutritional compositions, dietary or nutritional supplements, andmethods described herein can comprise, consist of, or consistessentially of the essential elements and limitations described herein,as well as any additional or optional ingredients, components, orlimitations described herein or otherwise useful in nutritional formulaapplications.

The present nutritional compositions, dietary supplements, and methodswill now be described more fully hereinafter with reference to theaccompanying drawings in which exemplary embodiments of the disclosureare shown. However, these compositions and methods may be embodied inmany different forms and should not be construed as limited to therepresentative embodiments set forth herein. The exemplary embodimentsare provided so that this disclosure will be both thorough and complete,and will fully convey the scope of the disclosure and enable one ofordinary skill in the art to make, use, and practice the teachings ofthe present disclosure.

Polyunsaturated fatty acids have numerous crucial biological functionsin mammals. For example, in humans, numerous polyunsaturated fatty acidshave been linked to cell membrane synthesis, metabolism, andmaintenance. Moreover, in adult humans, certain polyunsaturated fattyacids have been linked to potentially promoting improved cognitiveability and have been theorized as potential treatments for arthritis,cancer, diabetes, lupus, and psoriasis. Also, certain polyunsaturatedfatty acids have been linked to promoting retinal and brain developmentin fetuses and newborns.

For example, docosahexanoic acid (“DHA”) is one such polyunsaturatedfatty acid that has been implicated in playing a role in fetus andnewborn development as well as potentially playing a role in maintainingand/or promoting improved human cognitive ability. DHA is a long chainOmega-3 fatty acid derived from alpha-linolenic acid having double bondsat its 4, 7, 10, 13, 16, and 19 positions. In humans, DHA is primarilyfound in the brain's grey matter where it functions as a building blockfor membrane glycerophospholipids.

Although polyunsaturated fatty acids such as DHA are known to play animportant role in human cognitive ability, fetus development, andnewborn development, Western diets are often deficient in adequateamounts of these polyunsaturated fatty acids, and individuals oftenresort to polyunsaturated fatty acid supplementation to obtain adequatenutritional amounts of these polyunsaturated fatty acids to potentiallyavoid problems associated with polyunsaturated fatty acid deficiencies.For example, pregnant women and health conscious individuals oftensupplement their diets with fish oil and algae extracts in an attempt toobtain adequate amounts of polyunsaturated fatty acids. Moreover, infantformulas are often supplemented with polyunsaturated fatty acids derivedfrom either of these sources.

Although fish oil and algae extracts may be a source of polyunsaturatedfatty acids, numerous problems exist. For example, fish oil often has anundesirable smell and taste. Due to this problem, numerous fish oilformulations often include taste and smell masking agents. However, mosttaste and smell masking agents never completely mask the “fishy” tasteand smell of these oils. Moreover, these taste and smell masking agentsincrease production costs and the amount of time required to manufactureformulations having fish oil extracts. Thus, in many regards, fish oilsupplementation and production of supplements having these substancesremains undesirable due to these problems. Additionally, fish oil andalgal oil extracts do not have high quantities of DHA in thephospholipid form. And fish oil extracts also contain higher levels oftriglycerides and free fatty acids. The oxidation due to the higherlevels of triglycerides and free fatty acids are one of the reasons forthe undesirable smell and taste.

In addition, numerous problems exist for DHA derived from these sourcesand other commonly used forms of DHA. For example, DHA derived from fishoil and algae are primarily covalently bonded to triglycerides, andother types of pharmaceutical grade DHA are ethyl ester forms of DHA,which is not the bioactive form of DHA in the human body. In humans, DHAsupplementation in the form of DHA as an ethyl ester or free fatty acid,or triglycerides require multiple steps of metabolic processing to beuseful for certain biological functions. Thus, these forms of DHA areenergetically disfavored due to the amount of metabolic processingrequired by a subject's body.

Furthermore, when included in dietary supplements and formulations, theoxidation of DHA in ethyl ester or free fatty acid form, or triglycerideform leads to instability, decreased efficacy, decreased potency, andpotentially increased toxicity. The oxidation is also what contributesto the “fishy smell.” In addition to the problems mentioned above, thetriglyceride and ethyl ester forms of DHA often cause gastrointestinalproblems. Therefore, triglyceride and ethyl ester forms of DHA are oftenenterically coated in an attempt to reduce DHA oxidation andgastrointestinal problems associated with these forms of DHA, whichfurther increases production costs and the time required to manufacturedietary supplements having these substances.

DHA from algae or fish oxidizes (stinks) because it must be broken downto free fatty acid or ethyl ester DHA, which is then transported viaalbumin into the blood but then the supplement must provide high dosagesof DHA to “force” it or push it into blood-brain-barrier (“BBB”) or mustbe converted in the liver to lipid form DHA (only a small amount dothis). It takes a high dosages of docosahexanoic acid in free fatty acidor ethyl ester form and triglyceride form to cross the blood brainbarrier, which is why dietary supplements contain so much. Ultimately,the DHA must be metabolized into phospholipid-docosahexanoic acid(PL-DHA) in order to be circulating and then go into the BBB. PL-DHA isthe only form that can cross the BBB in significant numbers.

Therefore, it is an object of the disclosure to provide nutritionalcompositions and dietary supplements including a stable source of DHAthat avoids or reduces the problems associated with conventionalpolyunsaturated fatty acid supplementation. The forms of DHA that aremost beneficial for nutritional supplementation for cognitive functionsare in the phospholipid form, namely phosphatidylcholine docosahexanoicacid (“PC-DHA”), phosphatidylethanolamine docosahexanoic acid(“PE-DHA”), phosphatidylethanolamine-docosahexanoic acid (“PE-DHA”), andphosphatidylserine docosahexanoic acid (“PS-DHA”). These phospholipidsare conjugated to a docosahexanoic acid (PL-DHA), a tryglyceride withone other R group, where the R group contains a fatty acid.

Described herein are nutritional compositions and dietary supplementscontaining phospholipids and PL-DHA derived from eggs and methods ofmaking such nutritional compositions and dietary supplements. Thesenutritional compositions and dietary supplements can be administered toa subject to promote numerous health benefits, which include, but arenot limited to, promoting improved cognitive ability, retinal health,decrease inflammation, and neuronal health. Furthermore, thesenutritional compositions can be administered to pregnant women and/ornewborns to further promote brain and retinal development in fetuses andnewborn children.

Eggs contain many phospholipids and the amount of phospholipids in eggsmay be increased by supplementing the hens diet with polyunsaturatedfatty acids, particularly Omega-3 oils. Eggs contain various Omega-3fatty acids. Some of the phopholipids contained in eggs include:phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine,lysophosphatidyl choline, sphingomyelin, lysophosphtidyl ethanolamine,and combinations thereof. While fish, algae, and krill are commonsources for DHA supplementation, eggs is not a common source for DHAsupplementation due to the smaller amounts of DHA available compared tothe other sources.

Various phospholipids may be extracted from eggs. This “phospholipidextract” derived from eggs may also be referred to as the raw materialor ingredient for a dietary supplement. Reference to phospholipidextracts claimed by this disclosure will always be phospholipid extractsderived from eggs. The phospholipids contained in the phospholipidextract include phospholipid-docosahexanoic acid (PL-DHA) and otherphospholipids. Specifically, the PL-DHA of this disclosure may be PC-DHAand/or PE-DHA.

The phospholipid extract may also have some free fatty acids andtriglycerides, however, the phospholipid extract should preferably havea small amount triglycerides and free fatty acids. For the purposes ofthis disclosure, a “free fatty acid” is a fatty acid that is notcovalently bonded to a glycerol backbone or is an ethyl ester form ofDHA. A “triglyceride” lacks the phosphate group on the third carbon ofthe glycerol backbone, which also prevents choline, ethanolamine,serine, and inositol from binding to the triglyceride and making it aphospholipid.

The phospholipid extract or raw material may also phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, lysophosphatidyl choline,sphingomyelin, lysophosphtidyl ethanolamine, and combinations thereof.In one embodiment, the phospholipid extract has less than 5%triglycerides, in another embodiment, the phospholipid extract has lessthan 3% triglycerides. In one embodiment, the phospholipid extract hasless than 0.5% free fatty acids, in another embodiment, the phospholipidextract has less than 0.2% free fatty acids.

The phospholipid extract may sometimes be semi-solid and sticky. It canbe difficult to use in tablet or hard capsule products. The raw materialis often not soluble in oil, and using the material may be impracticalin use in softgel, as it may stick on the mill surface during grinding.

In certain aspects, these nutritional compositions and dietarysupplements include a body that encapsulates iron, folate, and anadditive, wherein the additive can include microparticles. In certainaspects, the microparticles have a substantially rounded shape andinclude a core. The core contains the raw material or phospholipidextract, which effectively and efficiently delivers the phospholipidextract to the subject. The core of the microparticle may be coated.These microparticles can be incorporated into the nutritionalcomposition to improve efficacy of phospholipid delivery.

In addition, these microparticles can be made by providing amicroparticle core made from a pharmaceutically acceptable material anda or phospholipid extract, then coating the microparticle core with afirst layer, and drying the coated microparticle core, thereby formingthe microparticle for the nutritional composition.

In other aspects the dietary supplement contains the phospholipidextract without the use of microparticles.

Additional features, aspects and advantages of the disclosure will beset forth in the detailed description which follows, and in part will bereadily apparent to those skilled in the art from that description orrecognized by practicing the teachings of the disclosure as describedherein. It is to be understood that both the foregoing generaldescription and the following detailed description present variousembodiments of the disclosure, and are intended to provide an overviewor framework for understanding the nature and character of thedisclosure as it is claimed. The accompanying drawings are included toprovide a further understanding of the disclosure, and are incorporatedin and constitute a part of this specification.

Nutritional Compositions

Described herein are nutritional compositions that include a bodyencapsulating iron, folate, and an additive, wherein the additiveincludes a microparticle having a core that includes a phospholipidextract and a coating. The body in the nutritional compositionsdescribed herein can include, but is not limited to, a tablet, anenteric coated tablet, a capsule, an enteric coated capsule, a softgelcapsule, or an enteric coated softgel capsule encapsulating iron,folate, and the additives, for example the microparticles, describedherein.

Docosahexanoic Acid (DHA)

The nutritional compositions and dietary supplements described hereininclude a pharmaceutically acceptable phospholipid-DHA derived from eggsand more preferably include organic DHA or DHA derived from organiceggs.

The docosahexanoic acid described herein generally includes thefollowing formula:

In certain aspects, the PL-DHA can include a phospholipid that ispreferably covalently bonded either directly to the DHA or indirectly(via a saturated or unsaturated lower alkyl linker having C₁-C₈) to theDHA. The phospholipid will have a glycerol backbone, an R₁ group on thefirst carboxyl group, and R₂ on the second carboxyl group, and aphosphate group on the third carboxyl group. For example, thephospholipid can include phosphatidylcholine, phosphatidylethanolamine,phosphatidylserine, or combinations thereof. In certain aspects, the R₁is a fatty acid. In certain aspects, the R₂ is a DHA.

The phosphate group of the phospholipid can include a primary aminegroup, a secondary amine group, a tertiary amine group, a trimethylamine group, or a combination thereof, and R₁ can include a hydrogen, ahydroxyl group, a saturated or unsaturated alkyl group, an alkoxy group,an omega-3 fatty acid, or any combination thereof. In certain aspects,the phosphate group will be bound to NH₃ ⁺ or N(CH₃)₃.

In certain aspects, PL-DHA of the nutritional composition may include1-palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine or1-hexadecanoyl-2-(4Z,7Z,10Z,13Z,16Z,19Z-docosahexaenoyl)-sn-glycero-3-phosphocholineor phosphatidylcholine-docosahexanoic acid (“PC-DHA”),1-stearoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine or1-octadecanoyl-2-(4Z,7Z,10Z,13Z,16Z,19Z-docosahexaenoyl)-sn-glycero-3-phosphoethanolamineor phosphatidylethanolamine-docosahexanoic acid (“PE-DHA”), orcombinations thereof. For example, Formula 1 depicts one representativeexample of PC-DHA.

The DHA shown in Formula 1 above is preferred over fish oil and algaederived forms of DHA for human nutrition because this DHA is in thephospholipid form, which aid in docosahexanoic stability and potency.Furthermore, these forms of DHA are more palatable to subjects thanthose derived from fish oil and algae extracts. Without wishing to bebound by theory, it is thought that these forms of DHA are more easilymetabolized by a subject and more easily cross the blood brain barrier.

FIG. 1 further illustrates this concept. For example, as shown in FIG.1, linolenic acid (i.e., “LNA”) and most forms of docosahexanoic acidare metabolized in the liver in order to cross the blood brain barrierbefore being used in various metabolic functions in the brain. Forexample, linolenic acid is converted into DHA, and DHA is further bondedto phosphatidylcholine to generate PC-DHA. After synthesizing PC-DHA,liver phospholipase A1 (PLase A1) hydrolyzes PC-DHA to form LysoPC-DHA,which can subsequently pass the blood brain barrier and can be used invarious metabolic functions in the brain such as maintaining neuronalmembrane glycerophospholipids and potentially preventing or reducingapoptosis of glial cells and neurons. As also shown in FIG. 1, if freePC-DHA is present, endothelial lipase can also hydrolyze this freePC-DHA to form LysoPC-DHA, which can also subsequently pass the bloodbrain barrier and be used in various metabolic functions in the brainsuch as maintaining neuronal membrane glycerophospholipids andpotentially preventing or reducing apoptosis glial cells and neurons.

FIG. 2 is an illustration of PC-DHA.

FIG. 3 is an illustration of PE-DHA.

The biochemical pathways that result in DHA conjugated to thephospholipid usually result in the DHA molecule binding to the numberone carboxyl group of the glycerol backbone. Though it is possiblehowever rare that the DHA may naturally be covalently bonded to thefirst carboxyl group. The many double carbon bonds of the DHA fatty acidcause the DHA fatty acid to be at an angle in relation to the glycerolbackbone, as opposed to a saturated fatty acid that will position itselfmore at a ninety degree angle to the glycerol backbone. Due to this tospatial configuration it is extremely rare for phopholipids to have twoDHA fatty acids conjugated to both the one carbon and the two carbonpositions, but it does occur in nature.

Krill oil is known to be a source of phospholipid and DHA nutritionalsupplementation, however, krill oil also contains significant amounts ofeicosapentaenoic acid (EPA).

Phospholipids extracts derived from eggs are also preferential in thategg yolk does not naturally contain eicosapentaenoic acid (EPA), butdoes contain arachidonic acid (AA). EPA interferes with the body's useof AA and probably does not belong in the mammalian body at all. Not allbiochemical pathways are perfect and some EPA may exist in nature inphospholipid extracts from eggs, however the amount would be small. Inone embodiment, the phospholipid extract comprises less than 1.5% EPA byweight. In one embodiment, the phospholipid extract comprises less than1% EPA by weight. In one embodiment, the phospholipid extract comprisesless than 0.5% EPA by weight.

DHA and AA are fatty acids that are needed by young children, pregnantand lactating women, and people with degenerative diseases involvingoxidative stress, especially those of the nervous system such asAlzheimer's.

When DHA is covalently linked to phosphatidylethanolamine, this form ofDHA can be easily converted to DHA forms that can cross the blood brainbarrier. For example, without wishing to be bound by theory, PE-DHA canbe converted into LysoPC-DHA by various metabolic processes andeventually cross the blood brain barrier.

In addition to crossing the blood brain barrier, PC-DHA and PE-DHA canbe used as a polyunsaturated fatty acid nutritional source that crossesthe placental barrier in pregnant mothers and provides a fetus withsufficient DHA amounts for adequate brain and retinal development.

In contrast to the PC-DHA and PE-DHA derived from eggs discussed above,DHA derived from fish oil and algae are primarily covalently bonded totriglycerides that do not include phosphate groups and/or phospholipids.Furthermore, the DHA derived from fish oil and algae often require farmore metabolic steps to be converted into a useful form of DHA that cancross the blood brain barrier and/or the placental barrier.

In certain aspects, the phospholipid extract includes PC-DHA and PE-DHA.In this aspect, the combination of PC-DHA and PE-DHA provided in amicroparticle or a dietary supplement can be 5% to 20% (by weight) ofthe overall amount of phospholipid extract in the dietary supplement. Inone aspect, PC-DHA can be 40% to 95% of the overall amount of PL-DHA inthe dietary supplement, and PE-DHA can be 5% to 60% of the overallamount of DHA in the dietary supplement. The PC-DHA and PE-DHA may alsobe used in a dietary supplement without the microencapsulation. In oneaspect, the at least 5% the phospholipid extract by weight is PC-DHA.And in another aspect, at least 5% the phospholipid extract by weight isPE-DHA. In one aspect, the at least 9% the phospholipid extract byweight is PC-DHA. And in another aspect, at least 9% the phospholipidextract by weight is PE-DHA.

Preferably the amount of triglycerides is low. In one aspect the amountof triglycerides is less than 5% of the phospholipid extract, and inanother aspect the amount of triglycerides is less than 3% of thephospholipid extract.

In certain aspects, the DHA described herein includes a ratio of PC-DHAto PE-DHA ranging from 20:1 to 1:1. In another embodiment, the DHAdescribed herein includes a ratio of PC-DHA to PE-DHA ranging from 5:1to 1:1.

DHA from Eggs

Eggs naturally contain a relatively high about of phospholipids andphospholipid conjugated DHA (PL-DHA). Additionally, to increase theamount of phospholipids and PL-DHA in eggs, chicken feed may besupplemented with a source of Omega-3 fatty acids like flax seeds, orother sources high in Omega-3 fatty acids. Between 200 mg to 300 mg ofusable phospholipid extract for nutritional supplementation can beextracted from one egg.

Omega-3 fatty acid enriched eggs have been shown to have 39% lessArachidonic Acid, an inflammatory Omega-6 fatty acid that most peopleeat too much of. It has also been shown that Omega-3 fatty acid enrichedeggs had 5 times as much Omega-3 fatty acid as conventional eggs.

Hens that are fed organic chicken feed potentially will be able toproduce DHA, including PC-DHA to PE-DHA, derived from eggs that may belabeled as a source of Organic DHA.

In certain aspects and as discussed further below, the PL-DHA is onlyincluded in the microparticles of the nutritional composition. In otheraspects, a portion of the phospholipid extract or PL-DHA can be includedin the microparticles and another portion of the phospholipid extract orPL-DHA can be included in a portion of the encapsulating body (e.g.,tablets, softgels, capsules, or chewable) that is not the microparticle.For example, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or 50% of theoverall amount of the phospholipid extract or PL-DHA can be in themicroparticles and the remainder of DHA can included in a portion of theencapsulating body that is not the microparticle.

The dietary supplement of the present disclosure may provide aphospholipid extract to be formulated with other vitamins and mineralsin the form of a tablet, softgel, capsule, chewable, or gummy. Thephospholipid extract may be provided in a dietary supplement in theamount ranging from 20 mg to 1500 mg, 100 mg to 900 mg, 100 mg to 800mg, 100 mg to 700 mg, 100 mg to 600 mg, 100 mg to 500 mg, 100 mg to 400mg, 100 mg to 300 mg, 100 mg to 200 mg, 200 mg to 800 mg, 200 mg to 650mg, 200 mg to 500 mg, 200 mg to 450 mg, 200 mg to 300 mg, 300 mg to 750mg, 300 mg to 600 mg, 300 mg to 500 mg, 300 mg to 450 mg, 300 mg to 400mg, 400 mg to 650 mg, 400 mg to 600 mg, 400 mg to 550 mg, 400 mg to 500mg, 400 mg to 450 mg, or any range having endpoints falling within anyof the preceding ranges. In one embodiments, the dietary supplementcontains 200 mg of the phospholipid extract.

Microparticles

Microparticles can be included in the disclosed nutritionalcompositions, and more specifically, the microparticles described hereinare preferably included within a tablet, an enteric coated tablet, acapsule, an enteric coated capsule, a softgel capsule, or an entericcoated softgel capsule.

The microparticles described herein generally include a microparticlecore and a layer or a plurality of layers coated on an outermost surfaceof the microparticle core. The microparticle core is preferably madefrom a pharmaceutically acceptable material that includes, but is notlimited to, tartaric acid, sugar, calcium carbonate, mannitol,microcrystalline cellulose, silica, starch or any combination thereof.These materials are mixed with the phospholipid extract. Thephospholipid extract may include PC-DHA, PE-DHA, or a mixture. PL-DHAand other Omega-3 fatty acids may also be incorporated in themicroparticle core.

It is preferable that the microparticle cores have a substantiallyrounded shape such that the microparticle cores do not aggregate duringthe production of the microparticles. In addition, because non-roundedshaped microparticle cores tend to aggregate and slow the productionprocess, it is preferable that the microparticle cores have a roundedshape to facilitate production of rounded microparticles.

The microparticle cores described herein can be generally produced withany known extrusion and spheronization techniques that can obtain thedesired microparticle diameters described below. For example, themicroparticle core material can be subjected to radial extrudingprocess, axial extruding process, cone extruding process, dome extrudingprocess, die roller extruding process, or basket extruding process.These materials can be generally extruded through a die or mesh havingvariable sizes. For example, the die and mesh sizes associated withthese extruding techniques can include 200 μm to 8000 μm, 300 μm to 7500μm, 300 μm to 4000 μm, 300 μm to 2000 μm, 400 μm to 2000 μm, 400 μm to1000 μm, and 500 μm to 800 μm to produce microparticle cores having adesired particle diameter. After extrusion, the microparticle corematerials can be further subjected to any known spheronization treatmentto further ensure sufficient microparticle core roundness.

In certain aspects, the microparticle cores can be further subjected toflow cytometry and separated based on desired microparticle coreroundness. For example, in certain aspects, the desired microparticlecore diameter ranges from 180 μm to 425 μm. In exemplary embodiments,the microparticles may have an accumulated volume average particlediameter D50 of from approximately 250 μm to approximately 425 μm tomeet 40-60 mesh requirements. Or the microparticles may have anaccumulated volume average particle diameter D70 of from approximately180 μm to approximately 250 μm to meet 60-80 mesh requirements. Or themicroparticles may have an accumulated volume average particle diameterD90 of from approximately 150 μm to approximately 180 μm to meet 80-100mesh requirements.

In an exemplary embodiment, the microparticle cores may have a volumeaverage particle size distribution index GSDv of approximately 1.30 orless. When GSDv is approximately 1.30 or less, little microparticle coreaggregation occurs and sufficient microparticle roundness can beobtained.

The accumulated volume average particle diameter D50 and the averageparticle size distribution index of the microparticle cores may bemeasured, for example, in the following manner. Based on a particle sizedistribution measured with such a measuring device as Coulter Counter TAII (available from Beckman Coulter, Inc.) or Multisizer II (availablefrom Beckman Coulter, Inc.), accumulated distributions of volume andnumber are each drawn from the small diameter side with respect to thedivided particle size ranges. The particle diameters where theaccumulated value is 16% are designated as volume D_(16V) and numberD_(16P), the particle diameters where the accumulated value is 50% aredesignated as volume D_(50V) and number D_(50P), and the particlediameters where the accumulated value is 84% are designated as volumeD_(84V) and number D_(84P). By using these values, the volume averageparticle size distribution index (GSDv) is calculated as(D_(84V)/D_(16V))^(1/2), and the number average particle sizedistribution index (GSDp) is calculated as (D_(84P)/D_(16P))^(1/2). Themicroparticles may have a shape factor SF1 of from approximately 110 toapproximately 140, and preferably from approximately 120 toapproximately 140, which results in microparticle cores having littleshape irregularity and having sufficient roundness (e.g., sphericalshape).

SF1 is a shape factor that shows the extent of unevenness on the surfaceof the microparticle cores, and is calculated as follows. An opticalmicrograph of the microparticle cores scattered on a glass slide isacquired to a Luzex image analyzer through a video cam, and SF1 iscalculated according to the following expression from the value obtainedby dividing square of the maximum length of the toner particles by theprojected area ((ML)²/A) for 50 toner particles, and the average valuethereof is designated as SF1.SF1=(ML)² /a×π/4×100wherein ML represents the maximum length of the toner particles, and Arepresents the projected area of the particles.

After obtaining microparticle cores having desirable roundness anddiameter, the microparticle cores are then subjected to a coatingprocess in which one or more coating layers are coated on an outermostsurface of the microparticle cores. For example, in certain aspects, anoutermost layer of the microparticle cores are coated with a solution,dispersion, or suspension. In some aspects, the coating may also containadditional DHA derived from eggs described above. In certain aspects,this coating can contain additional components that include, but are notlimited to tocopherol and other preservatives, stabilizers, andexcipients.

This coating can partially or completely coat the outermost layer of themicroparticle core. In certain aspects, it is preferable that thecoating completely coats the outermost layer of the microparticle core,and this combination of microparticle core and coating forms themicroparticle. Depending on the type of material used for themicroparticle core, it may be desirable to form an intermediary layerdisposed between the outermost surface of the microparticle core. In oneaspect the coating is pH dependant and may be used for targeting theabsorption at specific locations in the GI tract. In this aspect, theintermediary layer may reduce or prevent the docosahexanoic acid frominteracting and potentially blocking absorption.

In certain aspects, a plurality of coatings can be provided on themicroparticle core in order to form the desired microparticle. In thisaspect, the microparticle core can be coated with a coating containingphospholipid extract or PL-DHA on the outermost surface of themicroparticle core. In certain aspects, this coating is allowed to dryat a desired temperature and for a desired time period. Next, a secondcoating can be applied. In certain aspects, this second coating caninclude time release agents and additional excipients to more slowly andcontrollably administer, for example, the docosahexanoic acid derivedfrom eggs to a subject. These time release coatings are described ingreater detail further below. In further embodiments, additional coatinglayers can be provided on the microparticle cores.

In certain aspects, it is desirable that the microparticles do notexceed 500 μm in diameter because adverse effects such asgastrointestinal irritation may occur. It is also desirable that themicroparticles have a substantially uniform shape and particle diameterto ensure efficient delivery to the subject. For example, in certainaspects, the microparticles described herein are monodisperse and have apolydispersity index (PDI) ranging from about 1.5 to 1, from about 1.3to 1, and more preferably from about 1.2 to 1.

Organic DHA

The National Organic Program (NOP) under the direction of theAgricultural Marketing Service (AMS), an arm of the United StatesDepartment of Agriculture (USDA) is a national program that establishesnational standards for the production and handling of organicallyproduced products, including a National List of substances approved forand prohibited from use in organic production and handling. NOP iscodified in 7 CFR Part 205, which is incorporated by reference herein.

The final regulation declared that “Producers and handlers ofagricultural products used as ingredients in cosmetics, body careproducts, and dietary supplements could be certified as organicoperations. The ultimate labeling of cosmetics, body care products, anddietary supplements, however, has yet to be addressed.”

The USDA has stated that “There are agricultural products, includingpersonal care products, that, by virtue of their organic agriculturalproduct content, may meet the NOP standards and be labeled as “100percent organic,” “organic” or “made with organic” pursuant to the NOPregulations. Businesses that manufacture and distribute such productsmay be certified under the NOP, and such products may be labeled as “100percent organic,” “organic” or “made with organic” so long as they meetNOP requirements. Additionally, products that may be labeled “100percent organic” or “organic” may also carry the USDA organic seal.”

Except for exempt and excluded operations, each production or handlingoperation or specified portion of a production or handling operationthat produces or handles crops, livestock, livestock products, or otheragricultural products that are intended to be sold, labeled, orrepresented as “100 percent organic,” “organic,” or “made with organic(specified ingredients or food group(s))” must be certified. This meansthat the organic egg farm, the manufacturer of the organic eggphospholipid extract, the intermediate manufacturer, and the finalproduct manufacturer would need to be certified.

A certified operation must only use allowed substances, methods, andingredients for the production and handling of agricultural productsthat are sold, labeled, or represented as “100 percent organic,” or“organic,” for these products to be in compliance with the Act and theNOP regulations. Use of ionizing radiation, sewage sludge, and excludedmethods are prohibited in the production and handling of organicagricultural products.

The National List identifies synthetic substances, materials andingredients that may be used in organic farming and productionoperations. The list also highlights non-synthetic substances, materialsand ingredients that cannot be used. Notably, microcrystalline celluloseand methyl cellulose, which are ubiquitous excipients in dietarysupplements, are prohibited ingredients. This limitation clearly affectsfinished product manufacturers.

Most DHA and Omega fatty acids raw materials used in nutritional anddietary supplements are derived from algae and fish sources. Theextraction processes are not processes that are eligible to be certifiedas “organic.” These extraction techniques also tend to be responsiblefor the increased the amount of free fatty acids/ethyl ester DHA andtryglycerides as compared to the phospholipid extract of thisdisclosure. Additionally, some algae sources are genetically modifiedwhich also prevent products derived from the algae as certified“organic.” The present disclosure is the only known source of dietarysupplementation of DHA that is able to be certified as “organic” by theUSDA.

Organic egg production is the production of eggs through organic means.There are three main requirements for organic egg production:

-   -   Poultry can be exposed to antibiotics only during infectious        outbreak.    -   Poultry must be fed organic feed (no animal byproducts or        genetically-modified crops).    -   Poultry must have access to outdoors. It cannot be raised in        cages.

Organic production is also regulated by animal welfare audit system.Mistreatment of the chickens could potentially lead a farmer to losinghis organic certification.

Requirements of Phospholipid Extract Intermediate Manufacturers

Phospholipid extract intermediate manufacturers would need to becertified as organic operations.

(a) Mechanical or biological methods, including but not limited tocooking, baking, curing, heating, drying, mixing, grinding, churning,separating, distilling, extracting, slaughtering, cutting, fermenting,eviscerating, preserving, dehydrating, freezing, chilling, or otherwisemanufacturing, and the packaging, canning, jarring, or otherwiseenclosing food in a container may be used to process an organicallyproduced agricultural product for the purpose of retarding spoilage orotherwise preparing the agricultural product for market.(b) Nonagricultural substances allowed under §205.605 andnon-organically produced agricultural products allowed under §205.606may be used:

-   -   (1) In or on a processed agricultural product intended to be        sold, labeled, or represented as “organic,” pursuant to        §205.301(b), if not commercially available in organic form.    -   (2) In or on a processed agricultural product intended to be        sold, labeled, or represented as “made with organic (specified        ingredients or food group(s)),” pursuant to §205.301(c).        (c) The handler of an organic handling operation must not use in        or on agricultural products intended to be sold, labeled, or        represented as “100 percent organic,” “organic,” or “made with        organic (specified ingredients or food group(s)),” or in or on        any ingredients labeled as organic:    -   (1) Practices prohibited under paragraphs (e) and (f) of        §205.105.    -   (2) A volatile synthetic solvent or other synthetic processing        aid not allowed under §205.605: Except, That, nonorganic        ingredients in products labeled “made with organic (specified        ingredients or food group(s))” are not subject to this        requirement.

In one embodiment, this disclosure provides for phospholipids to be usedin a dietary supplement that may be labeled “organic” according to theUSDA guidelines. In one embodiment, this disclosure provide for DHA tobe used in a dietary supplement that may be labeled “organic” accordingto the USDA guidelines. In one embodiment, this disclosure provide forPL-DHA to be used in a dietary supplement that may be labeled “organic”according to the USDA guidelines.

Eggs may be farmed following the USDA guidelines for “organic” foods.The phospholipids from the eggs from organically raised and fed hens arethen extracted to be “Organic Phospholipid-DHA,” “Organic DHA.” or“Organic Phospholipids.” Then the organic phospholipid extract may beincluded in a dietary supplement in the form of a tablet, capsule,softgel, chewable tablet/softgel, or gummy.

Folic Acid and Folate

A pharmaceutically acceptable form of folic acid, folic acidderivatives, folate, reduced folate, or any combination thereof isincluded within the dietary supplement. Folate has been shown to play arole in nucleotide synthesis in mammals. Specifically, folic acid isknown to play a role in various methylation processes in humans and morespecifically in the synthesis of thymine from uracil (i.e., dUMP,deoxyuracil monophosphate). In adult humans, folic acid supplementationhas been implicated in reducing megaloblastic anaemia often associatedwith folate deficiency and/or side effects associated with variousmedical treatments (e.g., chemotherapy).

In addition, folic acid has been shown to play a role in preventingneural tube defects that occur during pregnancy. For example, althoughthe molecular and physiological mechanisms are currently unknown, folicacid supplementation has been theorized to reduce the occurrence ofspina bifida by up to 70%. Thus, for at least these reasons, it isdesirable to include a pharmaceutically acceptable form of folic acidand folic acid derivatives in the disclosed dietary supplements.

Various forms of folate are present in high concentrations in eggs. Forexample, folic acid, reduced folate, dihydrofolate, tetrahydrofolate,10-formylfolic acid, 5-formyltetrahydrofolate, and5-methyltetrahydrofolate are present in egg yolks. Therefore, in certainaspects and to potentially lower production costs of the discloseddietary supplements, it is desirable to extract folate from eggs whileobtaining the phospholipid extract.

In certain aspects, folate derivatives having substituents at its N⁵ orN¹⁰ position have increased stability and are less prone to cleavage andoxidation. Thus, these folic acid derivatives may be favored in thedisclosed dietary supplements. For example, in certain aspects, it ispreferable that at least one of folic acid, 10-formyl folic acid,5-formyl tetrahydrofolate, 5-methyl tetrahydrofolate, or any combinationthereof is present at a higher concentration in the nutritionalcomposition than dihydrofolate and tetrahydrofolate, and in certainaspects, dihydrofolate and/or tetrahydrofolate are not present in thedietary supplement.

The total biologically active amount of folate: folic acid andderivatives thereof present in the dietary supplement ranges from 100 μgto 15 mg. Total biologically active amounts of folic acid andderivatives thereof below 100 μg are potentially inadequate to preventor reduce problems associated with folate deficiencies (e.g.,megaloblastic anaemia, neural tube defects in fetuses, etc.). Pregnantmothers are recommended to take between 400 mcg and 1 mg of folate. Highdosages of folate have been shown to improve a number of human ailmentsand conditions including impaired cognitive function, ADHD, memory loss,diabetic peripheral neuropathy, and depression. Therefore, in certainaspects, the total biologically active amount of folate and derivativesthereof present in the dietary supplement from 100 μg to 15 mg, 400 μgto 800 μg, 400 μg to 1000 μg, 1 mg to 3 mg, 3 mg to 6 mg, 7.5 mg to 15mg, or any range having endpoints falling within any of the precedingranges.

In certain aspects, the total amount of folate, folic acid, and reducedfolate derivatives present in the nutritional composition ranges from0.1 mg to 20 mg based on the total weight of the nutritionalcomposition.

Folate may be provided through many different pharmaceuticallyacceptable salts

In certain aspects, the folic acid and folate derivatives are notincluded in the microparticles of the nutritional composition. In otheraspects, a portion of the folic acid and folate derivatives are includedin any of the coated layers of the microparticles and another portion ofthe folic acid and folate derivatives are included in the encapsulatingbody (e.g., tablets and capsules) that is not the microparticle. Forexample, in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%,55%, or 50% of the overall amount of the folic acid and folic acidderivatives are not in the microparticles, and in this aspect, theremainder of the folic acid and folic acid derivatives can included inthe microparticle.

Iron

A pharmaceutically acceptable form of iron is also included within thedisclosed Iron may be provided through many different pharmaceuticallyacceptable salts. In the human body, iron is a key component, which isoften complexed with various porphyrin rings to facilitate numerousmetabolic and biological processes. More specifically, iron is oftencomplexed with heme groups to form hemoglobin, and iron is also oftencomplexed with various cytochromes (e.g., Cytochrome C in themitochondria) to carry out electron transport during the production ofadenosine tri-phosphate (i.e., ATP) in the mitochondria.

In humans, iron deficiency is usually associated with various anemias,thrombocytosis, decreased immunity, increased susceptibility to sepsis,increased maternal mortality, and atrophy of mucous membranes (e.g.,Plummer-Vinson syndrome). Furthermore, iron deficiency in pregnantmothers, can potentially lead to low birth weights, preterm birth, andunder development in a newborn child. Thus, for at least these reasons,it is desirable to include a pharmaceutically acceptable form of iron inthe disclosed nutritional compositions.

In the disclosed nutritional composition, it is preferable to includeiron in the total amount of about 0.1 mg/kg to 6 mg/kg, 2.5 mg/kg to 5.5mg/kg, 3 mg/kg to 5 mg/kg, 3.5 mg/kg to 4.5 mg/kg, or any range havingendpoints falling within any of the preceding ranges of a subject's bodymass. For example, if the subject has a body mass of 100 kg (i.e., 220lbs), it is preferable to orally administer 10 mg to 600 mg of iron tothis subject when using the 2 mg/kg to 6 mg/kg range. Total amounts ofiron in the dietary supplement falling below 1 mg/kg are potentiallyinadequate to prevent or reduce problems associated with irondeficiency. Furthermore, total amounts of iron exceeding 6 mg/kg may beassociated with unwanted side effects associated such as iron poisoningif administered for an extended period of time. Therefore, in certainaspects, the total amount of iron thereof present in the dietarysupplement ranges from 10 mg to 600 mg, 10 mg to 500 mg, 10 mg to 400mg, 10 mg to 300 mg, 10 mg to 200 mg, 10 mg to 100 mg, 50 mg to 500 mg,50 mg to 400 mg, 50 mg to 300 mg, 50 mg to 200 mg, 50 mg to 100 mg, 100mg to 500 mg, 100 mg to 450 mg, 100 mg to 400 mg, 100 mg to 350 mg, 100mg to 300 mg, 100 mg to 250 mg, 100 mg to 200 mg, 100 mg to 150 mg, 200mg to 400 mg, 200 mg to 350 mg, 200 mg to 300 mg, 200 mg to 250 mg, 300mg to 500 mg, 300 mg to 450 mg, 300 mg to 400 mg, 300 mg to 350 mg, orany range having endpoints falling within any of the preceding ranges.

Iron may be provided through many different pharmaceutically acceptablesalts or chelates and one skilled in the art would know of these saltsand chelates. Some pharmaceutically acceptable forms of ironsupplementation may be through carbonyl, ferrous gluconate, ferrousfumerate, iron amino acid complexes, ferronyl carbonate, iron chelates,among others not listed.

In certain aspects, the total amount of Iron present in the dietarysupplement is about 27.5 mg. In certain aspects, the total amount ofiron present in the dietary supplement ranges from 0.5 mg to 100 mgbased on the total weight of the dietary supplement.

In certain aspects, iron is not included in the microparticles of thedietary supplement. In other aspects, a portion of iron is included inany of the coated layers of the microparticles and another portion ofiron is included in the encapsulating body (e.g., tablets and capsules)that is not the microparticle. For example, in certain aspects, at least90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or 50% of the overall amount ofiron is not in the microparticles, and in this aspect, the remainder ofthe iron can be included the microparticle.

Other Ingredients

In certain embodiments and as discussed further below, the discloseddietary supplement can optionally include pharmaceutically acceptableVitamin B₆, Vitamin B₁₂, N-Acetyl-L-Cysteine (NAC), Coenzyme Q10 (CoQ10) or any combination thereof.

In certain embodiments, the disclosed nutritional composition canoptionally include pharmaceutically acceptable Vitamin B₆. Vitamin B₆,and more specifically pyridoxal-5-phosphate, is involved in numerousbiological reactions occurring in a subject including amino acidmetabolism, hemoglobin synthesis, neurotransmitter synthesis, lipidmetabolism, and gluconeogenesis (e.g., glycogenolysis). Vitamin B₆ mayalso be supplemented as pyridoxine HCl. Other forms of Vitamin B₆ areknown and one skilled in the art would be aware of the various forms.Thus, to further complement the effects of DHA, folic acid andderivatives thereof, and iron mentioned above, Vitamin B₆ can be addedto the dietary supplement. In certain aspects, the total amount ofVitamin B₆ present in the dietary supplement ranges from 0.1 mg to 100mg, 0.1 mg to 75 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 0.1 mg to 10 mg,0.1 mg to 7.5 mg, 0.1 mg to 5 mg, 0.1 mg to 3 mg, 0.1 mg to 2.5 mg, 0.1mg to 2 mg, 0.1 mg to 1 mg, 0.1 mg to 0.5 mg, 0.2 mg to 10 mg, 0.2 mg to7.5 mg, 0.2 mg to 5 mg, 0.2 mg to 2.5 mg, 0.2 mg to 2.0 mg, 0.2 mg to1.5 mg, 0.2 mg to 1 mg, 0.3 mg to 10 mg, 0.3 mg to 7.5 mg, 0.3 mg to 5mg, 0.3 mg to 2.5 mg, 0.3 mg to 2.0 mg, 0.3 mg to 1.5 mg, 0.3 mg to 1mg, 0.5 mg to 5 mg, 0.5 mg to 4 mg, 0.5 mg to 3 mg, 0.5 mg to 2.5 mg,0.5 mg to 2.0 mg, 0.5 mg to 1.5 mg, 0.5 mg to 1 mg, 0.75 mg to 3 mg,0.75 mg to 2.5 mg, 0.75 mg to 2.0 mg, 0.75 mg to 1.5 mg, 0.75 mg to 1mg, 1 mg to 2 mg, 1 mg to 1.5 mg or any range having endpoints fallingwithin any of the preceding ranges.

In certain aspects, the total amount of Vitamin B₆ present in thedietary supplement ranges from 0.5 mg to 55 mg based on the total weightof the dietary supplement. In one aspect, the total amount of Vitamin B₆is 26 mg. In another aspect, the total amount of Vitamin B₆ is 35 mg. Inone aspect, the total amount of Vitamin B₆ is 25 mg.

In certain aspects, Vitamin B₆ is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Vitamin B₆is included in any of the coated layers of the microparticles andanother portion of Vitamin B₆ is included in the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Vitamin B₆ is not in the microparticles,and in this aspect, the remainder of Vitamin B₆ can be included in themicroparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Vitamin B₁₂. Vitamin B₁₂ plays arole in DNA synthesis, regulation of DNA synthesis, and fatty acidsynthesis. Vitamin B₁₂ has been implicated in playing a key role inmaintaining normal brain function and regulating the nervous system. Inhumans, Vitamin B₁₂ deficiency can cause severe and irreversible damageto the brain and nervous system, which may manifest with symptoms ofmania and psychosis. Thus, to potentially prevent Vitamin B₁₂ deficiencyand to further complement the effects of DHA, folic acid and derivativesthereof, and iron discussed above, Vitamin B₁₂ can be added to thedietary supplement. Vitamin B₁₂ may also be supplemented ascyanocobalamin, or preferably methylcobalamin. Other forms of VitaminB₁₂ are known and one skilled in the art would be aware of the variousforms.

In certain aspects, the total amount of Vitamin B₁₂ present in thedietary supplement ranges from 0.1 μg to 5 μg, 0.1 μg to 4 μg, 0.1 μg to3 μg, 0.1 μg to 2 μg, 0.1 μg to 1 μg, 0.3 μg to 4.5 μg, 0.3 μg to 3.5μg, 0.3 μg to 2.5 μg, 0.3 μg to 2 μg, 0.3 μg to 1 μg, 0.4 μg to 3.5 μg,0.4 μg to 3 μg, 0.4 μg to 2.5 μg, 0.4 μg to 2 μg, 0.4 μg to 1 μg, 0.5 μgto 3 μg, 0.5 μg to 2.5 μg, 0.5 μg to 2 μg, 0.5 μg to 2 μg, 0.5 μg to 0.9μg, or any range having endpoints falling within any of the precedingranges. In certain aspects, the total amount of Vitamin B₁₂ present inthe dietary supplement ranges from 10 μg to 5 mg based on the totalweight of the dietary supplement. In one aspect, the total amount ofVitamin B₁₂ is 12 mcg. In another aspect, the total amount of VitaminB₁₂ is 2 mg.

In certain aspects, Vitamin B₁₂ is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Vitamin B₁₂is included in any of the coated layers of the microparticles andanother portion of Vitamin B₁₂ is included in a portion of theencapsulating body (e.g., tablets and capsules) that is not themicroparticle. For example, in certain aspects, at least 90%, 85%, 80%,75%, 70%, 65%, 60%, 55%, or 50% of the overall amount of Vitamin B₁₂ isnot in the microparticles, and in this aspect, the remainder of VitaminB₁₂ can be included in the microparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable N-Acetyl-L-Cysteine in the range offrom 100 mg to 1000 mg, 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to700 mg, 100 mg to 600 mg, 100 mg to 500 mg, 100 mg to 400 mg, 100 mg to300 mg, 100 mg to 200 mg, 200 mg to 800 mg, 200 mg to 650 mg, 200 mg to500 mg, 200 mg to 450 mg, 200 mg to 300 mg, 300 mg to 750 mg, 300 mg to600 mg, 300 mg to 500 mg, 300 mg to 450 mg, 300 mg to 400 mg, 400 mg to650 mg, 400 mg to 600 mg, 400 mg to 550 mg, 400 mg to 500 mg, 400 mg to450 mg, or any range having endpoints falling within any of thepreceding ranges.

In certain aspects, the total amount of N-Acetyl-L-Cysteine present inthe dietary supplement ranges from 1 mg-1 g based on the total weight ofthe dietary supplement.

In certain aspects, N-Acetyl-L-Cysteine is not included in themicroparticles of the nutritional composition. In other aspects, aportion of N-Acetyl-L-Cysteine is included in any of the coated layersof the microparticles and another portion of N-Acetyl-L-Cysteine isincluded in a portion of the encapsulating body (e.g., tablets andcapsules) that is not the microparticle. For example, in certainaspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or 50% of theoverall amount of N-Acetyl-L-Cysteine is not in the microparticles, andin this aspect, the remainder of N-Acetyl-L-Cysteine can be included inthe microparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Coenzyme Q10 (CoQ 10). CoQ 10 is avitamin-like substance that is present primarily in mammalianmitochondria. In humans, CoQ10 has been implicated in oxidativephosphorylation and electron chain transport in the mitochondria, whichultimately results in the production of ATP. Furthermore, CoQ10 has anantioxidant function that reduces or prevents lipid peroxidation.Therefore, in certain aspects, CoQ10 may act synergistically with theDHA described herein as well as Vitamin B₆ and/or Vitamin B₁₂ to promotecognitive function by promoting lipid synthesis and lipid maintenanceoccurring, for example, in the brain while concurrently reducing lipiddamage occurring from lipid peroxidation.

In certain aspects, the total amount of CoQ10 present in the dietarysupplement ranges from 0.5 mg to 7 mg, 0.5 mg to 6 mg, 0.5 mg to 5 mg,0.5 mg to 4 mg, 0.5 mg to 3 mg, 0.5 mg to 2 mg, 0.5 mg to 1 mg, 1 mg to6.5 mg, 1 mg to 6 mg, 1 mg to 5.5 mg, 1 mg to 5 mg, 1 mg to 4.5 mg, 1 mgto 4 mg, 1 mg to 3.5 mg, 1 mg to 3 mg, 1 mg to 2.5 mg, 1 mg to 2 mg, 2mg to 6 mg, 2 mg to 5.5 mg, 2 mg to 5 mg, 2 mg to 4.5 mg, 2 mg to 4 mg,2 mg to 3.5 mg, 2 mg to 3 mg, 2 mg to 2.5 mg, 3 mg to 6 mg, 3 mg to 5.5mg, 3 mg to 5 mg, 3 mg to 4.5 mg, 3 mg to 4 mg, 3 mg to 3.5 mg, 4 mg to5.5 mg, 4 mg to 5 mg, 4 mg to 4.5 mg, or any range having endpointsfalling within any of the preceding ranges. In certain aspects, thetotal amount of CoQ10 present in the dietary supplement ranges from 50mg to 200 mg based on the total weight of the dietary supplement.

In certain aspects, CoQ10 is not included in the microparticles of thenutritional composition. In other aspects, a portion of CoQ10 isincluded in any of the coated layers of the microparticles and anotherportion of CoQ10 is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of CoQ10 is not in the microparticles, and inthis aspect, the remainder of CoQ10 can be included in a portion of theencapsulating body that is not the microparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Vitamin C in the range of from 10 mgto 1000 mg.

In certain aspects, the total amount of Vitamin C present in the dietarysupplement is about 70 mg.

In certain aspects, Vitamin C is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Vitamin C isincluded in any of the coated layers of the microparticles and anotherportion of Vitamin C is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Vitamin C is not in the microparticles, andin this aspect, the remainder of Vitamin C can be included in themicroparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Vitamin D in the range of from 50 IUto 2000 IU.

In certain aspects, the total amount of Vitamin D present in the dietarysupplement is about 1000 IU.

In certain aspects, Vitamin D is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Vitamin D isincluded in any of the coated layers of the microparticles and anotherportion of Vitamin D is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Vitamin D is not in the microparticles, andin this aspect, the remainder of Vitamin D can be included in themicroparticle.

In certain embodiments, the disclosed nutritional composition canoptionally include pharmaceutically acceptable Vitamin E in the range offrom 10 IU to 1000 IU.

In certain aspects, the total amount of Vitamin E present in thenutritional composition is about 10 IU.

In certain aspects, Vitamin E is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Vitamin E isincluded in any of the coated layers of the microparticles and anotherportion of Vitamin E is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Vitamin E is not in the microparticles, andin this aspect, the remainder of Vitamin E can be included in themicroparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Calcium in the range of from 5 mg to200 mg. Calcium may be provided through many different pharmaceuticallyacceptable salts or chelates and one skilled in the art would know ofthese salts or chelates.

In certain aspects, the total amount of Calcium present in the dietarysupplement is about 15 mg.

In certain aspects, Calcium is not included in the microparticles of thenutritional composition. In other aspects, a portion of Calcium isincluded in any of the coated layers of the microparticles and anotherportion of Calcium is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Calcium is not in the microparticles, andin this aspect, the remainder of Calcium can be included in themicroparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Iodine in the range of from 50 mcgto 300 mcg. Iodine may be provided through many differentpharmaceutically acceptable salts and one skilled in the art would knowof these salts. In one aspect, iodine is provided as potassium iodide.

In certain aspects, the total amount of Iodine present in the dietarysupplement is about 150 mcg.

In certain aspects, Iodine is not included in the microparticles of thenutritional composition. In other aspects, a portion of Iodine isincluded in any of the coated layers of the microparticles and anotherportion of Iodine is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Iodine is not in the microparticles, and inthis aspect, the remainder of Iodine can be included in themicroparticle.

In certain embodiments, the disclosed dietary supplement can optionallyinclude pharmaceutically acceptable Magnesium in the range of from 5 mgto 100 mg. Magnesium may be provided through many differentpharmaceutically acceptable salts and chelates and one skilled in theart would know of these salts and chelates. In one aspect, magnesium isprovided as magnesium oxide.

In certain aspects, the total amount of Magnesium present in the dietarysupplement is about 20 mg.

In certain aspects, Magnesium is not included in the microparticles ofthe nutritional composition. In other aspects, a portion of Magnesium isincluded in any of the coated layers of the microparticles and anotherportion of Magnesium is included in a portion of the encapsulating body(e.g., tablets and capsules) that is not the microparticle. For example,in certain aspects, at least 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or50% of the overall amount of Magnesium is not in the microparticles, andin this aspect, the remainder of Magnesium can be included in themicroparticle.

In addition, any standard pharmaceutically acceptable excipient can beused in the nutritional composition. For example, these excipients caninclude diluents (e.g., mannitol, sorbitol, lactose, sucrose, andcompressible sugars such as DiPac™ (dextrinized sucrose), available fromAustin Products Inc., Holmdel, N.J.), splitting or swelling agents(e.g., polyvinyl polypyrrolidone, croscarmellose sodium (e.g.,Ac-Di-Sol™ available from FMC BioPolymer, Philadelphia, Pa.), starchesand derivatives, cellulose and derivatives, microcrystalline celluloses,such as Avicel™ PH 101 or Avicel™ CE-15 (a microcrystalline modifiedwith guar gum), both available from FMC BioPolymer, Philadelphia, Pa.),lubricating agents (e.g., magnesium stearate), and flow agents (e.g.,colloidal silicon dioxide, such as Cab-O-Sil M5 available from CabotCorporation, Kokomo, Ind.).

The phospholipid extract as a raw material can be provided in a dietarysupplement as a natural product extracted from egg without anyadditional compounds. In some aspects the phospholipid extract may beblended with an antioxidant or other stabilizer for preservation;however, in another aspect, the phospholipid extract is not mixed withadditional excipients but is simply added to a dietary supplement. Ifthe phospholipid extract is not mixed with antioxidants, stabilizers, orexcipients, refrigerating the phospholipid extract will improvestability and shelf life.

Other DHA raw ingredients used for nutritional supplementation (sourcedfrom fish, algae, and krill) are mixed with many excipients and powdersand the actual DHA content is only about 10% of the raw ingredient byweight. Thus, 100 mg of some DHA powders only actually provide 10 mg ofDHA. The phospholipid extract provides more potent nutritionalsupplementation since a 100 mg amount of phospholipid extract wouldprovide 20 mg of DHA, and the rest of the supplement would contain otherphospholipids, which do provide nutritional benefit that the otherpowders would not.

Also, sweeteners can be included in the dietary supplement describedherein. For example, sweeteners can be used to impart a pleasant flavorto the composition. Suitable sweeteners for use in the presentdisclosure include natural sweeteners such as sucrose, dextrose,fructose, invert sugar, mannitol, sorbitol, and the like, as well assynthetic sweeteners such as saccharin, aspartame, acesulfame potassium,cyclamates, and other commercial artificial sweeteners well-known tothose of skill in the art. A preferred sweetener is acesulfame K(Sunett™ available from Nutrinova, Frankfort, Germany). The sweetener isadded in an amount to achieve a desired sweetness. Typically, thesweetener is present in an amount from about 1.0 wt % to about 5.0 wt %of the overall weight of the nutritional composition. Since the dietarysupplement may capitalize on the DHA being sourced from eggs,egg-friendly flavors are also preferred, which include vanilla and dulcede leche or caramel. Other flavors as chocolate or strawberry are alsoworkable. Those skilled in the part will appreciate that the amount ofsweetener may vary depending on the strength of the particular sweetenerused and the levels approved by the regulatory authorities for use inpharmaceutical products.

Time Release Coatings

In certain aspects, the outermost surface of the encapsulating body andthe outermost surface of the microparticle can independently include atime release coating. For example, in certain aspects the outermostsurface of the encapsulating body can include a time release coatingwhile such a coating is omitted from the microparticle. In otheraspects, the outermost surface of the microparticle can include a timerelease coating while such a coating is omitted from the outermostsurface of the encapsulating body.

Examples of these time release agents can include but are not limited tohydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxypropylmethyl cellulose phthalate, pullulan, gelatin, collagen, casein, agar,gum arabic, dextrin, ethyl cellulose, methyl cellulose, chitin,chitosan, mannan, carboxymethylethyl cellulose, sodium carboxymethylcellulose, polyethylene glycol, sodium alginate, poly(vinyl alcohol),cellulose acetate, poly(vinylpyrrolidone), silicone, poly(vinyl acetal)diethylamino acetate, albumin, adenine, cystine, D-tyrosine, or anycombination thereof.

In certain aspects, these time release coatings may range from 1-15% ofthe overall weight of the dietary supplement.

EXAMPLES Example 1 Microparticles Coated With Phospholipid Extract

Microparticle cores can be prepared by extrusion spheronizer technology,and the microparticle cores can be subsequently made of phospholipidextract, cellulose, starch, lactose, mannitol. PL-DHA may comprise 25%to 50% of the microparticle core.

A pharmaceutically acceptable source of phospholipid extract may bemixed with a pharmaceutically acceptable neutral material such astartaric acid, sugar sphere, calcium carbonate, mannitol,microcrystalline cellulose, silica, or starch which is then subjected toan extruding step to obtain microparticle cores in which 95% of themicroparticle cores have an average particle diameter ranging from 90 μmto 500 μm. These materials are mixed for 15-25 minutes and then extrudedout an extruder.

Next, the microparticle cores are placed into a spheroidizer at 500 rpmfor 5 to 10 second to ensure that sufficient microparticle coreroundness is obtained.

After ensuring proper microparticle core roundness has been obtained,the microparticle cores are organized into a bed of microparticle coresthat are subjected to a coating step. Next, the microparticle cores mayalso be (i) directly coated with an enteric coating to provide adissolution rate profile and/or (ii) directly coated with a protectivecoating: solution, suspension, or dispersion or. The enteric coating maybe L30D on different polymers. The enteric coating may comprise 1-5% ofthe weight of the total microparticle composition. The protectivecoating may be hpmc. The protective coating may comprise 1-10% of theweight of the total microparticle composition.

Example 2 Exemplary Nutritional Composition Formulations

Table 1 lists exemplary formulations of the dietary supplement of thepresent disclosure. The list is exemplary only and this disclosure isnot limited to the examples below.

TABLE 1 Exemplary Exemplary Exemplary Exemplary Formulation 1Formulation 2 Formulation 3 Formulation 4 Ingredient (wt %) (wt %) (wt%) (wt %) Total Egg Phospholipid 200 mg 90-95 mg 90-95 mg 90-95 mg(“Phospholipid extract”)t PC-DHA 20 mg 9 mg 9 mg 9 mg PE-DHA 5 mg 4.5 mg4.5 mg 4.5 mg Folic Acid or other folate 1 mg 3 mg 6 mg 15 mg (totalfolate) Iron 27.5 mg 0 mg 0 mg 0 mg Vitamin B6 (any form) 26 mg 35 mg 0mg 0 mg Vitamin E 20 IU 0 mg 0 mg 0 mg Vitamin B12 (any form) 12 mcg 2mg 2 mg 0 mg Calcium 15 mg 0 mg 0 mg 0 mg NAC (any form) 0 mg 0 mg 600mg 0 mg The formulations above do not include excipients, binders,stablizers, etc.

Example 3 Single Ingredient DHA Supplement

The dietary supplement of the present disclosure may provide aphopholipid extract formulated as a single ingredient provided in atablet, softgel, capsule, chewable, or gummy. The phospholipid extractmay be provided as a single ingredient dietary supplement in the rangesfrom 50 mg to 1000 mg, 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700mg, 100 mg to 600 mg, 100 mg to 500 mg, 100 mg to 400 mg, 100 mg to 300mg, 100 mg to 200 mg, 200 mg to 800 mg, 200 mg to 650 mg, 200 mg to 500mg, 200 mg to 450 mg, 200 mg to 300 mg, 300 mg to 750 mg, 300 mg to 600mg, 300 mg to 500 mg, 300 mg to 450 mg, 300 mg to 400 mg, 400 mg to 650mg, 400 mg to 600 mg, 400 mg to 550 mg, 400 mg to 500 mg, 400 mg to 450mg, or any range having endpoints falling within any of the precedingranges.

The foregoing description provides embodiments of the disclosure by wayof example only. It is envisioned that other embodiments may performsimilar functions and/or achieve similar results. Any and all suchequivalent embodiments and examples are within the scope of the presentdisclosure and are intended to be covered by the appended claims

It should be emphasized that the embodiments described herein are merelypossible examples of implementations, merely set forth for a clearunderstanding of the principles of the present disclosure. Manyvariations and modifications may be made to the described embodiment(s)without departing substantially from the spirit and principles of thepresent disclosure. Further, the scope of the present disclosure isintended to cover any and all combinations and sub-combinations of allelements, features, and aspects discussed above. All such modificationsand variations are intended to be included herein within the scope ofthe present disclosure, and all possible claims to individual aspects orcombinations of elements or steps are intended to be supported by thepresent disclosure.

One should note that conditional language, such as, among others, “can,”“could,” “might,” or “may,” unless specifically stated otherwise, orotherwise understood within the context as used, is generally intendedto convey that certain embodiments include, while alternativeembodiments do not include, certain features, elements and/or steps.Thus, such conditional language is not generally intended to imply thatfeatures, elements and/or steps are in any way required for one or moreparticular embodiments or that one or more particular embodimentsnecessarily include logic for deciding, with or without user input orprompting, whether these features, elements and/or steps are included orare to be performed in any particular embodiment. Unless statedotherwise, it should not be assumed that multiple features, embodiments,solutions, or elements address the same or related problems or needs.

Various implementations described in the present disclosure may includeadditional systems, methods, features, and advantages, which may notnecessarily be expressly disclosed herein but will be apparent to one ofordinary skill in the art upon examination of the following detaileddescription and accompanying drawings. It is intended that all suchsystems, methods, features, and advantages be included within thepresent disclosure and protected by the accompanying claims.

What is claimed is:
 1. A manufactured dietary supplement comprising: aphospholipid extract at a concentration ranging from 50 mg to 2 g, thephospholipid extract having phosphatidylcholine-docosahexanoic acid andphosphatidylethanolamine-docoshexanoic acid at a ratio ranging from 1:1to 2:1, wherein the amount of phosphatidylcholine-docosahexanoic acidcomprises from 7.5% to 10% the amount of phospholipids in thesupplement, and an effective amount of stabilizer that improves shelflife.
 2. The manufactured dietary supplement of claim 1, wherein thephospholipid extract is derived from eggs.
 3. The manufactured dietarysupplement of claim 1, wherein the phospholipid extract contains lessthan 5%, by weight, triglycerides.
 4. The manufactured dietarysupplement of claim 1, wherein the phospholipid extract contains lessthan 0.5%, by weight, free fatty acids.
 5. The manufactured dietarysupplement of claim 1, wherein the phospholipid extract is derived froma non-genetically modified organism.
 6. The manufactured dietarysupplement of claim 1, wherein the phospholipid extract the phospholipidextract comprises less than 0.5% eicosapentaenoic acid.
 7. Themanufactured dietary supplement of claim 1, wherein the phospholipidextract comprises an additional phospholipid that is not conjugated toDHA selected from phosphatidylcholine, phosphatidylethanolamine,lysophosphatidyl choline, sphingomyelin, lysophosphtidyl ethanolamine,and combinations thereof.
 8. The manufactured dietary supplement ofclaim 1, wherein the dietary supplement additionally comprises at leastone vitamin.
 9. The manufactured dietary supplement of claim 1, whereinthe phosphatidylcholine-docosahexanoic acid comprises 10%, by weight, ofthe phospholipid extract.
 10. The manufactured dietary supplement ofclaim 1, wherein the phosphatidylethanolamine-docosahexanoic acidcomprises from 5% to 10%, by weight, of the phospholipid extract. 11.The manufactured dietary supplement of claim 1, wherein from 15% to 20%of the phospholipid extract, by weight, is phospholipid-docosahexanoicacid.
 12. The manufactured dietary supplement of claim 1, wherein themanufactured dietary supplement is a tablet, softgel, capsule, orchewable.
 13. The manufactured dietary supplement of claim 1, whereinthe phospholipid extract additionally comprises arachidonic acid. 14.The manufactured dietary supplement of claim 1, wherein the phospholipidextract comprises at least 70% phosphatidylcholine.
 15. The manufactureddietary supplement of claim 1, wherein the stabilizer is a tocopherol.